An effective method of sonication-assisted Agrobacterium-mediated transformation of chickpeas

Abstract

An efficient and reproducible transformation method of sonication-assisted Agrobacteriummediated transformation (SAAT) was developed for chickpea (Cicer arietinum L.). Agrobacterium tumefaciens (LBA4404) harboring pCAMBIA1305.2 was used to transform decapitated embryo explants of two cultivars of chickpeas. By using a series of co-cultivation, callus induction, shoot initiation and root inducing media, a large number of transgenic plants were recovered. Transient expressions of GUS gene were detected by X-Gluc histochemical assay in transformed tissues. DNA analysis of T0 and T1 plants by PCR and Southern hybridization confirmed the integration of transgenes in initial and next generation transformants in different transgenic lines. The transformation efficiency was more than two times higher in SAAT treatment than simple Agrobacterium without sonication.

Physical wounding-assisted Agrobacterium-mediated transformation of juvenile cotyledons of a biodiesel-producing plant, Jatropha curcas L.

“…To this end, we attempted to enhance the transfer of DNA from Agrobacterium to plant cells by increasing the contact between Agrobacterium and the plant cells. A wounding treatment is crucial for efficient transformation and has been successfully employed to improve transformation efficiency in many plants (Du and Pijut 2009;Dutt et al 2007;de Oliveira et al 2009;Pathak and Hamzah 2008;Santarem et al 1998;Trick and Finer 1997;Weeks et al 2008). During wounding by sonication, the explosion of small bubbles from ultrasonic cavitation causes mechanical damage and creates micro-wounds on plant cells.…”

“…Abrasion of the apical node by sand-vortexing was used in alfalfa (Weeks et al 2008) and ultrasonic wounding has also been used to increase the transformation efficiency in many plant tissues (Pathak and Hamzah 2008;Santarem et al 1998;Trick and Finer 1997).…”

Highly Efficient <i>in Vitro</i> Regeneration Method of Endangered Medicinal Plant <i>Heliotropium kotschyi</i> (Ramram) in the Kingdom of Bahrain

“…Stem segments (Figure 1(B)) were surface sterilized following modified method [13], were washed in running tap water for fifteen minutes to remove dust particles from the surface of stem and leaf. Then stem segments were kept for five minutes successively in 1% of Lux solution, 0.5% of Mercric chloride solution with few drops of Tween 20, 0.1% of Copral solution, while the respective solutions were discarded in each step and were washed with autoclaved distilled water thoroughly (5 -6 times) in each step to remove the used solutions.…”

Effect of plant growth regulators on regeneration of the endangered medicinal plant Calligonum comosum L. Henry in the Kingdom of Bahrain

“…The stem segments were surface sterilized following modified method of Pathak and Hamzah (2008), in running tap water for 5 min to remove dust particles. Then the stem segments were kept for 5 min successively in 1% of lux solution, 0.5% of mercuric chloride solution with few drops of tween 20, 0.1% of copral solution, while the respective solutions were discarded in each step by thorough washing (5 to 6 times) with autoclaved water.…”